This assay employs an antibody specific for rat Leptin coated on a 96-well plate. Standards and samples are pipetted into the wells and Leptin present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-rat Leptin antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Leptin bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

FunctionMay function as part of a signaling pathway that acts to regulate the size of the body fat depot. An increase in the level of LEP may act directly or indirectly on the CNS to inhibit food intake and/or regulate energy expenditure as part of a homeostatic mechanism to maintain constancy of the adipose mass.

Involvement in diseaseDefects in LEP may be a cause of obesity (OBESITY) [MIM:601665]. It is a condition characterized by an increase of body weight beyond the limitation of skeletal and physical requirements, as the result of excessive accumulation of body fat.